Testosterone Parameter Assay Kit

Catalog # Availability Size / Price Qty
PKGE010
KGE010
SKGE010
Control Products Available
Multi-species Testosterone ELISA Standard Curve
2 Images
Product Details
Procedure
Citations (32)
FAQs
Supplemental Products
Reviews (1)

Testosterone Parameter Assay Kit Summary

Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (20 uL), EDTA Plasma (20 uL), Heparin Plasma (20 uL)
Sensitivity
0.041 ng/mL
Assay Range
0.0 - 10 ng/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Measures Testosterone levels in various samples
Cross-reactivity
Cross-reactivity observed with 1 or more available related molecules.Cross-species reactivity not tested.
Interference
Interference observed with 1 or more available related molecules.

Product Summary

The Parameter Testosterone kit is a 4.5 hour competitive enzyme immunoassay designed to measure Testosterone in cell culture supernates, serum, and plasma.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 1.25 3.5 5.8 1.26 3.67 6.03
Standard Deviation 0.05 0.11 0.17 0.07 0.23 0.41
CV% 4 3.1 2.9 5.6 6.3 6.8

Recovery

The recovery of Testosterone spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 103 95-117
EDTA Plasma (n=4) 94 85-107
Heparin Plasma (n=4) 92 84-101
Serum (n=4) 93 86-104

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Testosterone were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Multi-species Testosterone ELISA Linearity

Data Examples

Multi-species Testosterone ELISA Standard Curve

Product Datasheets

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Testosterone

Testosterone is an important secreted androgen synthesized from cholesterol-derived pregnenolone. Circulating testosterone is predominantly bound to proteins, including specific sex hormone binding globulin (SHBG) and nonspecific proteins such as albumin. It is believed that bioavailable testosterone includes the free steroid and the albumin-bound form. Testosterone is the main androgen secreted by the Leydig cells of the testes and affects both primary and secondary sexual development.

Alternate Names:
17b-Hydroxyandrost-4-ene-3-one; 17b-Testosterone ; Testosterone
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Primary Antibody Solution
  4.   Add 50 µL of Primary Antibody Solution to each well (except the non-specific binding (NSB) wells).
  5.   Incubate at room temperature for 1 hour on a horizontal orbital microplate shaker.
  6.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  7. Calibrator Diluent
  8.   Add 100 µL of Calibrator Diluent to the NSB wells.
  9.   Add 100 µL of Calibrator Diluent to the zero standard (B0) wells.

  10. 100 µL Standard, Control, or Sample
  11.   Add 100 µL of Standard, control, or sample to the remaining wells.

  12. 50 µL Conjugate
  13.   Add 50 µL of Conjugate to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours on the shaker.

  14. 200 µL Substrate Solution
  15. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  16. 50 µL Stop Solution
  17. Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Testosterone Parameter Assay Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

32 Citations: Showing 1 - 10
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  1. Development of Level A In Vitro-Vivo Correlation for Electrosprayed Microspheres Containing Leuprolide: Physicochemical, Pharmacokinetic, and Pharmacodynamic Evaluation
    Authors: DS Lee, DW Kang, GW Choi, HG Choi, HY Cho
    Pharmaceutics, 2020;12(1):.
    Species: Rat
    Sample Types: Plasma
  2. Resistance Exercise in a Hot Environment Alters Serum Markers in Untrained Males
    Authors: A Eskandari, M Fashi, A Saeidi, D Boullosa, I Laher, A Ben Abderr, G Jabbour, H Zouhal
    Front Physiol, 2020;11(0):597.
    Species: Human
    Sample Types: Serum
  3. Targeted Deletion of PTEN in Kisspeptin Cells Results in Brain Region- and Sex-Specific Effects on Kisspeptin Expression and Gonadotropin Release
    Authors: AL Negrón, G Yu, U Boehm, M Acosta-Mar
    Int J Mol Sci, 2020;21(6):.
    Species: Mouse
    Sample Types: Plasma
  4. Loss of p53 in mesenchymal stem cells promotes alteration of bone remodeling through negative regulation of osteoprotegerin
    Authors: T Velletri, Y Huang, Y Wang, Q Li, M Hu, N Xie, Q Yang, X Chen, Q Chen, P Shou, Y Gan, E Candi, AP Margherita, M Agostini, H Yang, G Melino, Y Shi, Y Wang
    Cell Death Differ., 2020;0(0):.
    Species: Mouse
    Sample Types: Serum
  5. Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells
    Authors: Y Bai, C Zhu, M Feng, H Wei, L Li, X Tian, Z Zhao, S Liu, N Ma, X Zhang, R Shi, C Fu, Z Wu, S Zhang
    Stem Cell Res Ther, 2018;9(1):200.
    Species: Porcine
    Sample Types: Cell Culture Supernates
  6. Effect of speed endurance training and reduced training volume on running economy and single muscle fiber adaptations in trained runners
    Authors: C Skovgaard, D Christians, PM Christense, NW Almquist, M Thomassen, J Bangsbo
    Physiol Rep, 2018;6(3):.
    Species: Human
    Sample Types: Plasma
  7. Serum cholesterol levels and tumor growth in a PTEN-null transgenic mouse model of prostate cancer
    Authors: EH Allott, EM Masko, AR Freedland, E Macias, K Pelton, KR Solomon, EA Mostaghel, GV Thomas, SV Pizzo, MR Freeman, SJ Freedland
    Prostate Cancer Prostatic Dis., 2018;0(0):.
    Species: Mouse
    Sample Types: Serum
  8. Impairment of learning and memory of mice offspring at puberty, young adulthood, and adulthood by low-dose Cd exposure during pregnancy and lactation via GABAAR ?5 and ? subunits
    Authors: Q Zhao, L Gao, Q Liu, Y Cao, Y He, A Hu, W Chen, J Cao, C Hu, L Li, F Tao
    Ecotoxicol. Environ. Saf., 2018;166(0):336-344.
    Species: Mouse
    Sample Types: Serum
  9. Pharmacokinetic-Pharmacodynamic Model for the Testosterone-Suppressive Effect of Leuprolide in Normal and Prostate Cancer Rats
    Authors: DS Lee, SJ Kim, GW Choi, YB Lee, HY Cho
    Molecules, 2018;23(4):.
    Species: Rat
    Sample Types: Plasma
  10. Testosterone represses urinary excretion of the alpha-tocopherol metabolite alpha-carboxymethylhydroxychroman in rats
    Authors: N Fujita, A Takenaka
    J. Nutr. Biochem., 2018;62(0):59-64.
    Species: Rat
    Sample Types: Plasma
  11. Reproductive Hormones and Their Receptors May Affect Lung Cancer
    Authors: M Dou, K Zhu, Z Fan, Y Zhang, X Chen, X Zhou, X Ding, L Li, Z Gu, M Guo, M Yan, X Deng, P Shen, S Wang
    Cell. Physiol. Biochem., 2017;44(4):1425-1434.
    Species: Mouse
    Sample Types: Serum
  12. Testosterone in advance age: a New Zealand longitudinal cohort study: Life and Living in Advanced Age (Te Pu?waitanga o Ng? Tapuwae Kia Ora Tonu)
    Authors: MJ Connolly, N Kerse, T Wilkinson, O Menzies, A Rolleston, YH Chong, JB Broad, SA Moyes, S Jatrana, R Teh
    BMJ Open, 2017;7(11):e016572.
    Species: Human
    Sample Types: Serum
  13. Transplantation of CD51(+) stem Leydig cells: A new strategy for the treatment of testosterone deficiency
    Authors: ZJ Zang, J Wang, Z Chen, Y Zhang, Y Gao, Z Su, Y Tuo, Y Liao, M Zhang, Q Yuan, C Deng, MH Jiang, AP Xiang
    Stem Cells, 2017;0(0):.
    Species: Mouse
    Sample Types: Serum
  14. Lysophosphatidic acid acyltransferase 3 tunes the membrane status of germ cells by incorporating docosahexaenoic acid during spermatogenesis
    Authors: Y Iizuka-His, D Hishikawa, J Sasaki, K Takubo, M Goto, K Nagata, H Nakanishi, H Shindou, T Okamura, C Ito, K Toshimori, T Sasaki, T Shimizu
    J. Biol. Chem., 2017;0(0):.
    Species: Mouse
    Sample Types: Serum
  15. Effects of Microcystis on Hypothalamic-Pituitary-Gonadal-Liver Axis in Nile Tilapia (Oreochromis niloticus)
    Authors: J Chen, S Meng, H Xu, Z Zhang, X Wu
    Bull Environ Contam Toxicol, 2017;0(0):.
  16. Effect of lipid metabolism on male fertility
    Authors: N Kim, H Nakamura, H Masaki, K Kumasawa, KI Hirano, T Kimura
    Biochem. Biophys. Res. Commun, 2017;0(0):.
    Species: Human
    Sample Types: Plasma
  17. Phospholipase C-related but Catalytically Inactive Proteins Regulate Ovarian Follicle Development
    Authors: M Matsuda, M Hirata
    J. Biol. Chem., 2017;0(0):.
    Species: Mouse
    Sample Types: Serum
  18. Effect of increased and maintained frequency of speed endurance training on performance and muscle adaptations in runners
    Authors: Jens Bangsbo
    J. Appl. Physiol., 2016;0(0):jap.00537.201.
    Species: Human
    Sample Types: Plasma
  19. Cutting edge: c-Kit signaling differentially regulates type 2 innate lymphoid cell accumulation and susceptibility to central nervous system demyelination in male and female SJL mice.
    Authors: Russi A, Walker-Caulfield M, Ebel M, Brown M
    J Immunol, 2015;194(12):5609-13.
    Species: Mouse
    Sample Types: Serum
  20. Diet-induced obesity in male C57BL/6 mice decreases fertility as a consequence of disrupted blood-testis barrier.
    Authors: Fan, Yong, Liu, Yue, Xue, Ke, Gu, Guobao, Fan, Weimin, Xu, Yali, Ding, Zhide
    PLoS ONE, 2015;10(4):e0120775.
    Species: Mouse
    Sample Types: Serum
  21. Osteopontin deletion prevents the development of obesity and hepatic steatosis via impaired adipose tissue matrix remodeling and reduced inflammation and fibrosis in adipose tissue and liver in mice.
    Authors: Lancha A, Rodriguez A, Catalan V, Becerril S, Sainz N, Ramirez B, Burrell M, Salvador J, Fruhbeck G, Gomez-Ambrosi J
    PLoS ONE, 2014;9(5):e98398.
  22. Insulin-like 3-induced rat preantral follicular growth is mediated by growth differentiation factor 9.
    Authors: Xue, Kai, Kim, Ji Young, Liu, Jia-yin, Tsang, Benjamin
    Endocrinology, 2014;155(1):156-67.
    Species: Rat
    Sample Types: Cell Culture Supernates
  23. Inhibitor of differentiation 4 (ID4) inactivation promotes de novo steroidogenesis and castration-resistant prostate cancer.
    Authors: Patel, Divya, Knowell, Ashley E, Korang-Yeboah, Maxwell, Sharma, Pankaj, Joshi, Jugal, Glymph, Shanora, Chinaranagari, Swathi, Nagappan, Peri, Palaniappan, Ravi, Bowen, Nathan J, Chaudhary, Jaideep
    Mol Endocrinol, 2014;28(8):1239-53.
    Species: Human
    Sample Types: Cell Culture Supernates
  24. Alterations of bone density, microstructure, and strength of the distal radius in male patients with rheumatoid arthritis: a case-control study with HR-pQCT.
    Authors: Zhu T, Griffith J, Qin L, Hung V, Fong T, Au S, Li M, Lam Y, Wong C, Kwok A, Leung P, Li E, Tam L
    J Bone Miner Res, 2014;29(9):2118-29.
    Species: Human
    Sample Types: Serum
  25. Inhibitor of DNA binding 4 is expressed selectively by single spermatogonia in the male germline and regulates the self-renewal of spermatogonial stem cells in mice.
    Authors: Oatley MJ, Kaucher AV, Racicot KE
    Biol. Reprod., 2011;85(2):347-56.
    Species: Mouse
    Sample Types: Serum
  26. Testosterone regulates cell proliferation in aggressive fibromatosis (desmoid tumour).
    Authors: Hong H, Nadesan P, Poon R, Alman BA
    Br. J. Cancer, 2011;104(9):1452-8.
    Species: Mouse
    Sample Types: Serum
  27. Host-derived adiponectin is tumor-suppressive and a novel therapeutic target for multiple myeloma and the associated bone disease.
    Authors: Fowler JA, Lwin ST, Drake MT
    Blood, 2011;118(22):5872-82.
    Species: Human
    Sample Types: Serum
  28. Endocrine responses during overnight recovery from exercise: impact of nutrition and relationships with muscle protein synthesis.
    Authors: Betts JA, Beelen M, Stokes KA, Saris W HM, van Loon L JC
    Int J Sport Nutr Exerc Metab, 2011;21(5):398-409.
    Species: Human
    Sample Types: Plasma
  29. Skeletal muscle myofibrillar and sarcoplasmic protein synthesis rates are affected differently by altitude-induced hypoxia in native lowlanders.
    Authors: Holm L, Haslund ML, Robach P, van Hall G, Calbet JA, Saltin B, Lundby C
    PLoS ONE, 2010;5(12):e15606.
    Species: Human
    Sample Types: Plasma
  30. Castration Differentially Alters Basal and Leucine-Stimulated Tissue Protein Synthesis in Skeletal Muscle and Adipose Tissue.
    Authors: Jiao Q, Pruznak AM, Huber D, Vary TC, Lang CH
    Am. J. Physiol. Endocrinol. Metab., 2009;0(0):.
    Species: Rat
    Sample Types: Plasma
  31. Inhibition of tumor growth progression by antiandrogens and mTOR inhibitor in a Pten-deficient mouse model of prostate cancer.
    Authors: Zhang W, Zhu J, Efferson CL, Ware C, Tammam J, Angagaw M, Laskey J, Bettano KA, Kasibhatla S, Reilly JF, Sur C, Majumder PK
    Cancer Res., 2009;69(18):7466-72.
    Species: Mouse
    Sample Types: Plasma
  32. Antagonistic effects of a mixture of low-dose nonylphenol and di-n-butyl phthalate (monobutyl phthalate) on the Sertoli cells and serum reproductive hormones in prepubertal male rats in vitro and in vivo.
    Authors: Hu, Yang, Wang, Ruoyu, Xiang, Zou, Qian, Weiping, Han, Xiaodong, Li, Dongmei
    PLoS ONE, 0;9(3):e93425.
    Species: Rat
    Sample Types: Serum

FAQs

  1. Does R&D Systems® Testosterone Parameter Assay Kit, Catalog # KGE010, detect free or total testosterone?

    • 'Free' testosterone is typically defined as testosterone which is bioavailable, and not bound to sex hormone binding globulin (SHBG). Interference testing performed by spiking SHBG into testosterone-containing samples, demonstrates interference in testosterone detection with Catalog # KGE010, when the concentration of SHBG is greater than 100 ng/ml. (For further details on interference testing, please refer to the "Specificity" section in the product insert.) 

      Assays for 'total' testosterone measure both 'free' testostrone as well as testosterone bound to SHBG. Since KGE010 does not detect testosterone bound to SHBG, this assay does not measure 'total' testosterone.

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Testosterone Parameter Assay Kit
By Anonymous on 03/22/2018
Application: Sample Tested: Serum-free Cell Culture Media,Serum